5 replies to this topic

[moljul]

after trypsinization of cells, medium should be added to inactivate the enzymatic digestion!!

which compound in the medium is responsible for this inactivation?? is it the serum or is it other compound??


if it is the serum, could i inactivate also with medium containing less than 10% serum? my medium has 1% serum!!

thx for recommendation

[laurequillo]

As far as I know is the BSA in the serum.

I dont know about the 1% serum medium. If you are not sure you can just inactivate with "normal" medium  and then centrifuge and remove and add the 1%serum medium.

[klinmed]

moljul, on Jul 9 2010, 12:06 PM, said:

after trypsinization of cells, medium should be added to inactivate the enzymatic digestion!!

which compound in the medium is responsible for this inactivation?? is it the serum or is it other compound??


if it is the serum, could i inactivate also with medium containing less than 10% serum? my medium has 1% serum!!

thx for recommendation

The enzyme is inactivated by protease inhibitors present in the serum supplement. These include alpha-1-antitrypsin and alpha-2-macroglobulin. 1% FCS is enough especially if the bulk of the trypsin/EDTA is removed by centrifugation/resuspension of the cells.

Hope this helps.

[bob1]

You can also buy soybean trypsin inhibitor from many companies.  I have used it from Sigma.

[laurequillo]

So it looks like I was wrong. Good to know!

[Inmost sun]

laurequillo, on Jul 12 2010, 07:28 AM, said:

So it looks like I was wrong. Good to know!

I think you are not wrong since albumine and other bulk proteins in serum are targeted by trypsin and compete with cell surface proteins of cells